Simultaneous Determination of Saponins in Radix Glycyrrhizae, Notoginseng and Ginseng by High Performance Liquid Chromatography

To establish a method for determining five saponins(notoginsenoside R1, ginsenoside Rg1, ginsenoside Re, ginsenoside Rb1 and ammonium glycyrrhizinate) in Glycyrrhizae, Notoginseng and Ginseng, the high performance liquid chromatography with diode array detector(HPLC-DAD) method was applied to an Inertsil ODS-SP column(4.6 mm×250 mm, 5 μm) with a mobile phase consisting of acetonitrile-0.05% phosphoric acid in a gradient elution manner. The flow rate was 1.0 mL/min. The column temperature was 30 ℃ and the detection wavelengths were 203 nm and 237 nm, respectively. The linear ranges were 0.700,0—7.000,0 μg for R1(r=1.000,0), 0.751,1— 7.511,4 μg for Rg1(r=1.000,0), 0.677,2—6.771,6 μg for Re(r=1.000,0), 0.733,9—7.339,1 μg for Rb1(r= 1.000,0), and 0.540,0—5.399,8 μg for ammonium glycyrrhizinate(r=0.999,9), respectively. In addition, their average recoveries were 100.28%, 105.83%, 104.09%, 99.36% and 98.54%, respectively. The relative standard deviations(RSDs) of precision, reproducibility and recovery were all less than 1.5%. The results indicate that the method is simple, accurate and reproducible so that it can be used for the simultaneous determination of the five saponins in Chinese patent medicines containing the three kinds of herbs.

Phytochemical Analysis of Ziziphus jujuba Leaves in Six Cultivars at the Whole Life Stage by High Performance Liquid Chromatography

Leaves of Ziziphus jujuba Mill.（Z. jujuba） were used as a folk medicine to treat children suffering from typhoid fever, furuncle and ecthyma in China. The present study was to establish a high performance liquid chroma- tography-diode array detector（HPLC-DAD） method for quality control of Z. jujuba leaves. The validated method was applied to the simultaneous characterization and quantification of sixteen main constituents in thirty samples, which comprised the whole life stage of Z. ]ujuba leaves from six cultivars in Yulin, Shannxi Province, China. HPLC fin- gerprint file of Z. jujuba leaves was set up and the results indicate that the major constituents of Z. jujuba leaves are （-）-catechin（1）, quercetin-3-O-robinobioside（3）, rutin（4） and quercetin-3-O-a-L-arabinosyl-（1---~2）-a-L-rhamnoside（6） with the total contents of the nine flavonoids in respective batch ranging from 18.52 mg/g to 39.51 mg/g. In addition, the contents of compounds depend on both the season and the cultivar. During ripening of Z. jujuba leaves, flavo- noids of all the cultivars present a similar trend： initially decrease from 25th, Jun. to 5th, Aug., 2015, and then in- crease for a period of time, finally slightly fluctuate on 5th Oct. The highest contents of total flavonoids were regis- tered on 25th Jun., 2015. Different cultivars have 50%--90% variation on the content for the three classes of com- pounds. Taken together, the results demonstrate that Z. jujuba leaves have great potential to be a new health-promoting resource based on flavonoids and the best sampling time of them is in June.

Extraction optimization of coumarins from radix angelicae pubescentis by HPLC-DAD coupled with uniform design

Uniform design was used to optimize extraction condition of direct refluence extraction of coumarins from the Chinese traditional medicine of radix angelicae pubescentis(Duhuo); the sum peak area of coumarins separated with high performance liquid chromatography(HPLC) at detection wavelength of 320nm was considered as detection index, two factors of solvent concentration and extraction time were mainly studied at extraction temperature of 85℃ and a volume ratio of solvent to sample of 10∶1. Optimal subclass, quadric polynomial step by step aggression and neural network method were applied to process the experimental results. The results show that the first and second methods give the same factors combination (concentration of ethanol: 95%, extraction time: 3.6 h) and the second method is much better than the first one. The extraction model is consequently developed.

Study on chromatographic fingerprint of sarcandra glabra (Thunb.) by microwave-assisted extraction coupled to HPLC/DAD

Microwave-assisted extraction(MAE)was used for extraction of effective components of sarcandra glabra(Thunb.),and then chromatographic fingerprint of sarcandra glabra(Thunb.)was studied by high performance liquid chromatography/diode array detector(HPLC/DAD).The conditions of MAE were optimized by an orthogonal experiment,and then the authentication and validation of the chromatographic fingerprint were conducted.Nine peaks were identified as common peaks in the fingerprint chromatograms,and isofraxidin was considered as a reference compound and quantified.Relative standard deviations of retention time and peak area of each component were less than 3% and 8%,respectively.Similarity and difference analysis were conducted by use of PCA and relation coefficient.Twenty batches of sarcandra glabra(Thunb.)samples from two different producing areas could be classified into two different groups in the PCA model.The results showed that MAE-HPLC/DAD method was simple,efficient and stable for the study of complex chromatographic fingerprint of sarcandra glabra(Thunb.),which could provide more reliable and precise information for quality evaluation.

Simultaneous and interference-free determination of eleven nonsteroidal anti-inflammatory drugs illegally added into Chinese patent drugs using chemometrics-assisted HPLC-DAD strategy

In this work, a smart strategy that combines three-way high performance liquid chromatography-diode array detection(HPLCDAD) data with second-order calibration method based on alternating trilinear decomposition(ATLD) algorithm was proposed for simultaneous determination of eleven non-steroidal anti-inflammatory drugs(NSAIDs) illegally added into Chinese patent drugs and health products. All target analytes were rapidly eluted out within 14.5 min under a simple gradient elution. With the aid of the prominent "second-order advantage" of the ATLD algorithm, three HPLC problems, i.e. peak overlaps, unknown interferences and baseline drift, could be mathematically calibrated, and pure signals of target analytes could be extracted out from heavy-interference but information-rich HPLC-DAD data. The average spiked recoveries for all target analytes were in the range of 95.9%–106.4% with standard deviations lower than 7.5%. Validation parameters including sensitivity(SEN), selectivity(SEL), limit of detection(LOD), limit of quantitation(LOQ) and precisions of intra-day and inter-day were calculated to validate the accuracy of the proposed method, quantitative results were further confirmed by the classic HPLC method, which proved that chemometrics-assisted HPLC-DAD analytical strategy was highly efficient, accurate and green for drug-abuse monitoring of NSAIDs in Chinese patent drugs and health products.

Simultaneous Analysis of Indapamide and Related Impurities in Sustained-Release Tablets by a Single-Run HPLC-PDA Method

The contents of indapamide and related impurities in generic indapamide sustained-release tablets were simultaneously detected by a single-run high performance liquid chromatography equipped with photodiode array detector(HPLC-PDA)method for the quality control in this paper.The results showed the method had a good selectivity and was validated through linearity,limits of detection and quantification,recovery,and precision.The linear ranges of indapamide,2-methyl-1-nitroso-2,3-dihydro-1H-indole(impurity A,ImA),4-chloro-N-(2-methyl-1H-indol-1-yl)-3-sulphamoyl-benzamide(impurity B,ImB)and 4-chloro-3-sulfamoylbenzoic acid(impurity 1,Im1)were 0.028-1.80μg/mL(R=0.99995),0.060-1.20μg/mL(R=0.9996),0.0324-1.20μg/mL(R=0.99985)and 0.060-1.20μg/mL(R=0.9997)with detection limits of 0.0093,0.012,0.012 and 0.006μg/mL,respectively.ImA and Im1 were not detectable in the generic drug.The content of indapamide was 96.7%of the labeled amount with a relative standard deviation(RSD)of 1.30%,and the percentage of ImB relative to the labeled amounts of indapamide was 0.106%with an RSD of 1.82%.The content of other unspecified impurities all met the reference quality standards.The results provided references for the quality control and the quality standard study of generic indapamide sustained-release tablets.

Quantitative Analysis and Chemical Fingerprint Similarity for Quality Control of the Seeds of Paeonia suffruticosa Andr. by HPLC

According to the General Office of the State Council of China, tree peony seeds（TPSs, Paeonia suffruticosa Andr.） are considered an emerging source for edible oil. In Chinese folk medicine, TPSs are used for alleviating waist and leg pain. In the present study, a simple, accurate and rapid fingerprint method based on high performance liquid chromatography with diode array detection（HPLe-DAD） was developed and validated to quantify the profiles of ten representative compounds in TPSs. All standard calibration curves exhibited good linearity（R2〉0.9995） in the I-IPLC-DAD analysis. The recovery of the standards ranged from 96.37% to 102.10%. The results show that stilbe- noids are the major constituents in TPSs, and their total content ranges from 25.71 mg/g to 54.03 mg/g. Suffruticosol A, suffruticosol B, trans-e-viniferm, ampelopsin E and paeoniflorin are the major components in methanol- water（80：20, volume ratio） extracts with average contents of 3.962, 12.264, 5.826, 14.060 and 12.755 rag/g, respec- tively. Additionally, 14 batches of seeds collected from different regions were used to establish a reference fingerprint Their similarity values were higher than 0.944 except for the sample（0.863） from Linxia. These results collectively indicate that the quantitative HPLC fingerprint method can serve as a prerequisite quality control for TPSs, and is a promising resource for developing new herbal or food products.